An improved pollen number counting method using a cell counter and mesh columns
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(2020) 16:124 Kakui et al. Plant Methods https://doi.org/10.1186/s13007-020-00668-4
Open Access
METHODOLOGY
An improved pollen number counting method using a cell counter and mesh columns Hiroyuki Kakui1* , Eriko Tsurisaki2, Hidenori Sassa3 and Yoshinari Moriguchi1
Abstract Background: The determination of pollen number is important in evolutionary, agricultural, and medical studies. Tree species of the Cupressaceae family cause serious pollinosis worldwide. Although Japanese cedar (Cryptomeria japonica) is the most important forestry species in Japan, it is also the biggest cause of pollinosis in the country. Japanese cedar trees have been selected for growth speed and superior morphological traits and then cloned. These clones may vary in their pollen production, but there has been little research on how many pollen grains are produced by a single male strobilus (flower). A recently reported method for counting pollen number with a cell counter was applicable to Arabidopsis species and wheat, but was not suitable for Japanese cedar because the strobilus does not open with heating (e.g. 60 °C, overnight). Results: Here, we report an improved pollen counting method for Japanese cedar using a precise and rapid cell counter in combination with home-made mesh columns. The male strobilus was gently crushed using a pestle. Large and small debris were then removed using 100- and 20-μm mesh columns, respectively. We successfully detected pollen sizes and numbers that differed between two clones using this method. Conclusions: This improved method is not only suitable for counting pollen from Japanese cedar, but could also be applied to other species of the Cupressaceae family with hard scale tissue covering the pollen. Moreover, this method could be applied to a broader range of plant species, such as wheat, because there is no need to wait for anthesis and debris can be removed efficiently. Keywords: Pollen number, Pollen size, CASY cell counter, Japanese cedar (Cryptomeria japonica) Background The determination of pollen grain number is important in evolutionary, agricultural, and medical studies. From an evolutionary perspective, selfing plant species tend to produce lower pollen numbers than closely related outcrossing plant species [1–5]. The reduced pollen number in selfing plants is thought to decrease the cost of pollen production. From an agricultural perspective, domesticated species such as rice tend to have low pollen numbers [6]; however, the production of large numbers of pollen grains is one of the desired traits in hybrid wheat *Correspondence: [email protected]‑u.ac.jp 1 Graduate School of Science and Technology, Niigata University, Niigata, Niigata 950‑2181, Japan Full list of author information is available at the end of the article
breeding [7]. A high pollen number is also a desired trait for crops that require artificial pollination because artificial pollination require a lot of pollen [8, 9]. From a medical perspective, pollen is relevant because it can lead to an allergic reaction calle
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